The goal of this project is to develop a set of significantly improved cytogenetics specimen preparation and automated image analysis techniques for diagnosing leukemia and monitoring the progress, response to therapy and remission of the disease. These techniques will work on interphase cells, avoiding the need for culturing metaphase cells. This will reduce the time and cost requirements of leukemia diagnosis and therapy monitoring and provide more quantitative data for diagnosis and follow-up. Specifically, we will develop bone marrow specimen preparation, staining and labelling techniques and automated image analysis hardware and software to fractionate and quantify the numbers of interphase cells based on (1) cytogenetically normal versus abnormal, (2) proliferation index and (3) state of cell maturity. We will use (1) DNA probes on interphase nuclei to determine which cells have specific leukemia-related chromosomal abnormalities, (2) Anti-BRDU to determine proliferation index, and (3) computerized morphological image analysis to evaluate state of cell maturity. The analysis will be done on an automated microscope that can process 60 slides unattended. This new capability will significantly increase the ability of clinicians to diagnose leukemia and monitor therapy.